Repair of rhodopsin mRNA by spliceosome-mediated RNA trans-splicing: a new approach for autosomal dominant retinitis pigmentosa.
Détails
ID Serval
serval:BIB_23C134DDA937
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Repair of rhodopsin mRNA by spliceosome-mediated RNA trans-splicing: a new approach for autosomal dominant retinitis pigmentosa.
Périodique
Molecular therapy
ISSN
1525-0024 (Electronic)
ISSN-L
1525-0016
Statut éditorial
Publié
Date de publication
05/2015
Peer-reviewed
Oui
Volume
23
Numéro
5
Pages
918-930
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Publication Status: ppublish
Résumé
The promising clinical results obtained for ocular gene therapy in recent years have paved the way for gene supplementation to treat recessively inherited forms of retinal degeneration. The situation is more complex for dominant mutations, as the toxic mutant gene product must be removed. We used spliceosome-mediated RNA trans-splicing as a strategy for repairing the transcript of the rhodopsin gene, the gene most frequently mutated in autosomal dominant retinitis pigmentosa. We tested 17 different molecules targeting the pre-mRNA intron 1, by transient transfection of HEK-293T cells, with subsequent trans-splicing quantification at the transcript level. We found that the targeting of some parts of the intron promoted trans-splicing more efficiently than the targeting of other areas, and that trans-splicing rate could be increased by modifying the replacement sequence. We then developed cell lines stably expressing the rhodopsin gene, for the assessment of phenotypic criteria relevant to the pathogenesis of retinitis pigmentosa. Using this model, we showed that trans-splicing restored the correct localization of the protein to the plasma membrane. Finally, we tested our best candidate by AAV gene transfer in a mouse model of retinitis pigmentosa that expresses a mutant allele of the human rhodopsin gene, and demonstrated the feasibility of trans-splicing in vivo. This work paves the way for trans-splicing gene therapy to treat retinitis pigmentosa due to rhodopsin gene mutation and, more generally, for the treatment of genetic diseases with dominant transmission.
Mots-clé
Animals, Binding Sites, Cell Line, Transformed, Gene Expression, Genes, Dominant, Genetic Therapy, Genetic Vectors/genetics, Humans, Introns, Mice, Mice, Transgenic, Phenotype, Photoreceptor Cells/metabolism, Protein Interaction Domains and Motifs/genetics, Protein Transport, RNA Precursors/genetics, RNA Splicing, RNA, Messenger/genetics, Retinitis Pigmentosa/genetics, Retinitis Pigmentosa/therapy, Rhodopsin/chemistry, Rhodopsin/genetics, Rhodopsin/metabolism, Trans-Splicing, Transduction, Genetic
Pubmed
Web of science
Open Access
Oui
Création de la notice
15/07/2020 11:08
Dernière modification de la notice
16/07/2020 9:43